In epithelial tissues the hemidesmosome is considered to play an important role in epithelial-connective tissue adherence. Its reassembly during wound healing in epithelia appears essential for the establishment of firm attachment of epithelial cells to the underlying connective tissue. The hemidesmosome acts as the structural link between components of the extracellular matrix in the connective tissue and the cytoskeleton of the basal epithelial cell. The hemidesmosomal plaque is the anchorage site of bundles of keratin containing intermediate filaments along the basal surface of the epithelial cell. It is the goal of Dr. Jones to investigate the molecular structure and assembly of the hemidesmosome. He is able to undertake such analyses since he has derived a number of antibody preparations directed against the hemidesmosome and also molecular probes for certain hemidesmosomal plaque elements during the previous grant period. In addition, he recently identified a cell line, termed 804G, which readily assembles hemidesmosomes in culture.
The specific aims are: (1) He will continue studies of the isolation of hemidesmosomal polypeptides using a combination of chromatographic and immunopurification procedures. (2) He will monitor hemidesmosome assembly at the ultrastructural level in his recently described epithelial explant wound healing model. (3) He will clone and sequence hemidesmosomal components. The deduced amino acid sequences and secondary structural predictions that are derived from such analyses may indicate homologies with known polypeptides and may also suggest functions for hemidesmosomal polypeptides. (4) He will monitor hemidesmosome assembly in 804G cells at the morphological, biochemical and molecular levels. (5) He will investigate hemidesmosomal protein turnover in 804G cells. (6) He will identify glycosylated hemidesmosomal elements involved in the initiation of hemidesmosomal assembly and/or hemidesmosome-substratum adherence by adhesion assays. This will involve the use of monoclonal and polyclonal antibodies generated against both tissue hemidesmosomal components and the hemidesmosomes of 804G cells. (7) He will use transfection techniques to identify sequences of hemidesmosomal proteins necessary for the incorporation of elements into the hemidesmosome and assembly of the hemidesmosome complex.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM038470-05
Application #
3294904
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1988-04-01
Project End
1995-03-31
Budget Start
1992-04-01
Budget End
1993-03-31
Support Year
5
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Northwestern University at Chicago
Department
Type
Schools of Dentistry
DUNS #
005436803
City
Chicago
State
IL
Country
United States
Zip Code
60611
DeHart, Gregory W; Jones, Jonathan C R (2004) Myosin-mediated cytoskeleton contraction and Rho GTPases regulate laminin-5 matrix assembly. Cell Motil Cytoskeleton 57:107-17
deHart, Gregory W; Healy, Kevin E; Jones, Jonathan C R (2003) The role of alpha3beta1 integrin in determining the supramolecular organization of laminin-5 in the extracellular matrix of keratinocytes. Exp Cell Res 283:67-79
Gonzales, M; Weksler, B; Tsuruta, D et al. (2001) Structure and function of a vimentin-associated matrix adhesion in endothelial cells. Mol Biol Cell 12:85-100
Goldfinger, L E; Jiang, L; Hopkinson, S B et al. (2000) Spatial regulation and activity modulation of plasmin by high affinity binding to the G domain of the alpha 3 subunit of laminin-5. J Biol Chem 275:34887-93
Hopkinson, S B; Jones, J C (2000) The N terminus of the transmembrane protein BP180 interacts with the N-terminal domain of BP230, thereby mediating keratin cytoskeleton anchorage to the cell surface at the site of the hemidesmosome. Mol Biol Cell 11:277-86
Goldfinger, L E; Hopkinson, S B; deHart, G W et al. (1999) The alpha3 laminin subunit, alpha6beta4 and alpha3beta1 integrin coordinately regulate wound healing in cultured epithelial cells and in the skin. J Cell Sci 112 ( Pt 16):2615-29
Gonzales, M; Haan, K; Baker, S E et al. (1999) A cell signal pathway involving laminin-5, alpha3beta1 integrin, and mitogen-activated protein kinase can regulate epithelial cell proliferation. Mol Biol Cell 10:259-70
Chen, M; Marinkovich, M P; Jones, J C et al. (1999) NC1 domain of type VII collagen binds to the beta3 chain of laminin 5 via a unique subdomain within the fibronectin-like repeats. J Invest Dermatol 112:177-83
Hopkinson, S B; Findlay, K; deHart, G W et al. (1998) Interaction of BP180 (type XVII collagen) and alpha6 integrin is necessary for stabilization of hemidesmosome structure. J Invest Dermatol 111:1015-22
El-Ghannam, A; Starr, L; Jones, J (1998) Laminin-5 coating enhances epithelial cell attachment, spreading, and hemidesmosome assembly on Ti-6A1-4V implant material in vitro. J Biomed Mater Res 41:30-40

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